MCC950-CP-456773-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEMCC950Cat.No.:HY-12815CASNo.:210826-40-7Synonyms:CP-456773;CRID3分?式:C??H??N?O?S分?量:404.48作?靶點:NOD-likeReceptor(NLR)作?通路:Immunology/Inflammation儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗H2O:50mg/mL(123.62mM;Needultrasonic)DMSO:≥28mg/mL(69.22mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.4723mL12.3616mL24.7231mL5mM0.4945mL2.4723mL4.9446mL10mM0.2472mL1.2362mL2.4723mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；?旦配成溶液，請分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲存時，請在6個?內(nèi)使?，-20°C儲存時，請在1個?內(nèi)使?。體內(nèi)實驗請根據(jù)您的實驗動物和給藥?式選擇適當(dāng)?shù)娜芙?案。以下溶解?案都請先按照InVitro?式配制澄的儲備液，再依次添加助溶劑：(為保證實驗結(jié)果的可靠性，澄的儲備液可以根據(jù)儲存條件，適當(dāng)保存；體內(nèi)實驗的?作液，建議您現(xiàn)?現(xiàn)配，當(dāng)天使?；以下溶劑前顯?的百分?指該溶劑在您配制終溶液中的體積占?；如在配制過程中出現(xiàn)沉淀1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE、析出現(xiàn)象，可以通過加熱和/或超聲的?式助溶)1.請依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(6.18mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(6.18mM);Clearsolution3.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(6.18mM);ClearsolutionBIOLOGICALACTIVITY?物活性MCC950(CP-456773;CRID3)?種有效，選擇性的NLRP3抑制劑，在BMDMs和HMDMs中的IC50分別為7.5nM和8.1nM。IC50&TargetIC50:7.5nM(NLRP3,inBMDMs),8.1nM(NLRP3,inHMDMs)[1]體外研究MCC950blockscanonicalandnon-canonicalNLRP3activationatnanomolarconcentrations.MCC950specificallyinhibitsNLRP3butnotAIM2,NLRC4orNLRP1activation.TheeffectofMCC950onNLRP3inflammasomeactivationistestedinmousebonemarrowderivedmacrophages(BMDM)andhumanmonocytederivedmacrophages(HMDM).TheIC50ofMCC950inBMDMisapproximately7.5nM,whileinHMDMithasasimilarinhibitorycapacity(IC50=8.1nM).MCC950alsodosedependentlyinhibitIL-1βbutnotTNF-αsecretion.MCC950specificallyblockscaspase-11-directedNLRP3activationandIL-1βsecretionuponstimulationofthenon-canonicalpathway.NLRC4-stimulatedIL-1βandTNF-αsecretion(asactivatedbySalmonellatyphimurium)arenotinhibitedbyMCC950evenataconcentrationof10μM.MCC950doesnotinhibitcaspase-1activationorIL-1βprocessinginresponsetoS.typhimurium.Theexpressionofpro-caspase-1andpro-IL-1βincelllysatesisnotsubstantiallyaffectedbyMCC950treatment[1].體內(nèi)研究MCC950reducesInterleukin-1p(IL-1β)productionandattenuatestheseverityofexperimentalautoimmuneencephalomyelitis(EAE),adiseasemodelofmultiplesclerosis.Pre-treatmentwithMCC950reducesserumconcentrationsofIL-1βandIL-6whileitdoesnotconsiderablydecreasetheamountofTNF-α.TreatmentofmicewithMCC950delaystheonsetandreducedtheseverityofEAE.IntracellularcytokinestainingandFACSanalysisofbrainmononuclearcellsfrommicesacrificedonday22showsmodestlyreducedfrequenciesofIL-17andIFN-γproducingCD3+TcellsinMCC950treatedmiceincomparisonwithPBS-treatedmice.IFN-γandparticularlyIL-17producingcellnumbersarealsoreducedinboththeCD4+andγδ+sub-populationsofCD3+Tcells[1].PROTOCOLCellAssay[1]BMDMareseededat5×105/mLor1×106/mL,HMDMat5×105/mLandPBMCat2×106/mLor5×106/mLin96wellplates.Thefollowingdaytheovernightmediumisreplacedandcellsarestimulatedwith10ng/mLLPSfromEscherichiacoliserotypeEH100(ra)TLRgradfor3h.Mediumisremovedandreplacedwithserumfreemedium(SFM)containingDMSO(1:1,000),MCC950(0.001-10μM),Parthenolide(10μM)orBayercysteinylleukotrienereceptorantagonist1-(5-carboxy-2{3-[4-(3-2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEcyclohexylpropoxy)phenyl]propoxy}benzoyl)piperidine-4-carboxylicacid(40μM)for30min.Cellsarethenstimulatedwithinflammasomeactivators:5mMadenosine5’-triphosphatedisodiumsalthydrate(ATP)(1h),1μg/mLPoly(deoxyadenylic-thymidylic)acidsodiumsalt(PolydA:dT)transfectedwithLipofectamine200(3-4h),200μg/mLMSU(overnight)and10μMnigericin(1h)orS.typhimuriumUK-1strain.Cellsarealsostimulatedwith25μg/mLPolyadenylic-polyuridylicacid(4h).Fornon-canonicalinflammasomeactivationcellsareprimedwith100ng/mLPam3CSK4for4h,mediumisremovedandreplacedwithSFMcontainingDMSOorMCC950and2μg/mLLPSistransfectedusing0.25%FuGENEfor16h.SupernatantsareremovedandanalysedusingELISAkits.LDHreleaseismeasuredusingtheCytoTox96non-radioactivecytotoxicityassay[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]Administration[1]C57BL/6miceareimmunizedsubcutaneouslywith150μgofMOGpeptide35-55emulsifiedinCFAcontaining4mg/mL(0.4.mg/mouse)ofheat-killedMTB.Miceareinjectedi.p.with500ngpertussistoxin(PT:kaketsuken)ondays0and2.MCC950isadministeredi.p.tomice(10mg/kg)atinductionofthedisease,day0,1and2andevery2daysthereafter.Controlmiceareadministeredvehicle(PBS)atthesametimepoints.Miceareobservedforclinicalsignsofdiseasedaily(unblinded).Diseaseseverityisscoredasfollows:noclinicalsigns,0;limptail,1;ataxicgait,2;hindlimbweakness,3;hindlimbparalysis,4;andtetraparalysis,5.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?Science.2020Dec4;3